Targeted nanomedicines for the treatment of bone disease and regeneration.

Targeted delivery by either passive or active targeting of therapeutics to the bone is an attractive treatment for various bone related diseases such as osteoporosis, osteosarcoma, multiple myeloma, and metastatic bone tumors. Engineering novel drug delivery carriers can increase therapeutic efficacy and minimize the risk of side effects. Developmnet of nanocarrier delivery systems is an interesting field of ongoing studies with opportunities to provide more effective therapies. In addition, preclinical nanomedicine research can open new opportunities for preclinical bone-targeted drug delivery; nevertheless, further research is needed to progress these therapies towards clinical applications. In the present review, the latest advancements in targeting moieties and nanocarrier drug delivery systems for the treatment of bone diseases are summarized. We also review the regeneration capability and effective delivery of nanomedicines for orthopedic applications.

A sustainable and self-healable silk fibroin nanocomposite with antibacterial and drug eluting properties for 3D printed wound dressings.

The development of self-healable and 3D printable hydrogels with decent biocompatibility, mechanical durability, adhesiveness to tissues, and antibacterial activity is of great importance for wound healing applications. In this study, we present a sustainable and environmentally friendly composite hydrogel consisting of silk fibroin (SF), oxidized salep (OS), and kappa carrageenan nanoparticles (NPs) for efficient wound care. The injectable nanocomposite hydrogel is highly stretchable and exhibits strong tissue adhesiveness and self-healing response through Schiff-base cross-linking between OS and SF. The tunable shear-thinning viscoelastic properties of the hydrogel facilitate 3D bioprinting with excellent shape adaptability (97.7 ± 1.1% recovery), enabling the fabrication of complex-shaped constructs. In vitro release kinetics of tetracycline (TC) encapsulated in kappa carrageenan NPs indicate a distinctive Korsmeyer-Peppas profile, including an initial burst release followed by a triphasic pattern controlled by the embedded NPs within the hydrogel matrix. The composite hydrogel shows a remarkable broad-spectrum antibacterial activity with substantial zones of inhibition against S. aureus (34.00 ± 1.00 mm) and E. coli (27.60 ± 2.08 mm) after 24 h of incubation at 37 °C. The addition of TC further enhances the zones of inhibition by approximately 45% for S. aureus and 27% for E. coli. The control group without kappa NP incorporation shows no zone of inhibition, underscoring the critical role of the nanoparticles in imparting antibacterial activity to the hydrogel. Cytocompatibility assays show the high viability of fibroblast (L929) cells (>90%) in vitro. In vivo biocompatibility studies through subcutaneous implantation also do not show malignancy, infection, abscess, necrosis, epidermal or dermal modifications, or inflammation of the wounds after 14 days post-injection. H&E staining shows that the biodegradation of the developed hydrogel facilitates the growth of non-inflammatory cells, leading to the substitution of the injected hydrogel with autologous tissue. The detailed analyses affirm that the multifunctional injectable hydrogel with self-healing and antibacterial properties has high potential for wound healing and skin tissue engineering.

Trilayer composite scaffold for urethral reconstruction:in vitroevaluation of mechanical, biological, and angiogenic properties.

Regeneration of damaged urethral tissue remains a major challenge in the field of lower urinary tract reconstruction. To address this issue, various synthetic and natural biodegradable biomaterials are currently being explored for the fabrication of scaffolds that promote urethral regeneration and healing. In this study, we present an approach to fabricate a trilayer hybrid scaffold comprising a central layer of poly(lactic acid) (PLA) between two layers of chitosan. The chitosan/PLA/chitosan (CPC) scaffolds were fabricated by a sequential electrospinning process and their properties were evaluated for their suitability for urethral tissue engineering. The physical and biological properties of the CPC scaffolds were evaluated in comparison to electrospun PLA scaffolds and acellular dermis (Alloderm) as controls for a synthetic and a natural scaffold, respectively. Compared to the controls, the CPC scaffolds exhibited higher elastic modulus and ultimate tensile strength, while maintaining extensibility and suture retention strength appropriate for clinical use. The CPC scaffolds displayed significant hydrophilicity, which was associated with a higher water absorption capacity of the chitosan nanofibres. The degradation products of the CPC scaffolds did not exhibit cytotoxicity and promoted wound closure by fibroblastsin vitro. In addition, CPC scaffolds showed increased growth of smooth muscle cells, an essential component for functional regeneration of urethral tissue. Furthermore, in a chicken embryo-based assay, CPC scaffolds demonstrated significantly higher angiogenic potential, indicating their ability to promote vascularisation, a crucial aspect for successful urethral reconstruction. Overall, these results suggest that CPC hybrid scaffolds containing both natural and synthetic components offer significant advantages over conventional acellular or synthetic materials alone. CPC scaffolds show promise as potential candidates for further research into the reconstruction of the urethrain vivo.

Engineering Photo-Cross-Linkable MXene-Based Hydrogels: Durable Conductive Biomaterials for Electroactive Tissues and Interfaces.

Light-cured conductive hydrogels have attracted immense interest in the regeneration of electroactive tissues and bioelectronic interfaces. Despite the unique properties of MXene (MX), its light-blocking effect in the range of 300-600 nm hinders the efficient cross-linking of photocurable hydrogels. In this study, we investigated the photo-cross-linking process of MX-gelatin methacrylate (GelMa) composites with different types of photoinitiators and MX concentrations to prepare biocompatible, injectable, conductive, and photocurable composite hydrogels. The examined photoinitiators were Eosin Y, Irgacure 2959 (Type I), and lithium phenyl-2,4,6-trimethylbenzoyl phosphinate (Type II). The light-blocking effect of MX strongly affected the thickness, pore structure, swelling ratio, degradation, and mechanical properties of the light-cured hydrogels. Uniform distribution of MX in the hydrogel matrix was achieved at concentrations up to 0.04 wt % but the film thickness and curing times varied depending on the type of photoinitiator. It was feasible to prepare thin films (0.5 mm) by employing Type I photoinitiators under a relatively long light irradiation (4-5 min) while thick films with centimeter sizes could be rapidly cured by using Type II photoinitiator (<60 s). The mechanical properties, including elastic modulus, toughness, and stress to break for the Type II hydrogels were significantly superior (up to 300%) to those of Type I hydrogels depending on the MX concentration. The swelling ratio was also remarkably higher (648-1274%). A conductivity of about 1 mS/cm was attained at 0.1 mg/mL MX for the composite hydrogel cured by the Type I photoinitiator. In vitro cytocompatibility assays determined that the hydrogels promoted cell viability, metabolic activity, and robust proliferation of C2C12 myoblasts, which indicated their potential to support muscle cell growth during myogenesis. The developed photocurable GelMa-MX hydrogels have the potential to serve as bioactive and conductive scaffolds to modulate cellular functions and for tissue-device interfacing.

Biomimetic nanoengineered scaffold for enhanced full-thickness cutaneous wound healing.

Wound healing is a complex process based on the coordinated signaling molecules and dynamic interactions between the engineered scaffold and newly formed tissue. So far, most of the engineered scaffolds used for the healing of full-thickness skin wounds do not mimic the natural extracellular matrix (ECM) complexity and therefore are not able to provide an appropriate niche for endogenous tissue regeneration [1]. To address this gap and to accelerate the wound healing process, we present biomimetic bilayer scaffolds compositing of gelatin nanofibers (GFS) and photocrosslinkable composite hydrogels loaded with epidermal growth factors (EGF). The nanofibers operate as the dermis layer, and EGF-loaded composite hydrogels acted as the epidermis matrix for the full-thickness wound healing application. The hydrogels are composed of gelatin metacryloyl (GelMA) modified with silicate nanoplatelets (Laponite). To overcome the challenges of transdermal delivery of EGF, including short half-life and lack of efficient formulation precise, controlled delivery was attained by immobilization of EGF on Laponite. It is shown that the addition of 1wt% silicate nanoplatelet increases the compressive modulus of the hydrogels by 170%. In vitro wound closure analysis also demonstrated improved adhesion of the scaffolds to the native tissue by 3.5 folds. Moreover, the tunable hemostatic ability of the scaffolds due to the negatively charged nanoplatelets is shown. In an established excisional full-thickness wound model, an enhanced wound closure (up to 93.1 ± 1.5%) after 14 days relative to controls (GFS and saline-treated groups) is demonstrated. The engineered adhesive and hemostatic scaffolds with sustained release of the growth factors have the potential to stimulate complete skin regeneration for full-thickness wound healing.

Nanoengineered shear-thinning and bioprintable hydrogel as a versatile platform for biomedical applications.

The development of bioinks based on shear-thinning and self-healing hydrogels has recently attracted significant attention for constructing complex three-dimensional physiological microenvironments. For extrusion-based bioprinting, it is challenging to provide high structural reliability and resolution of printed structures while protecting cells from shear forces during printing. Herein, we present shear-thinning and printable hydrogels based on silicate nanomaterials, laponite (LA), and glycosaminoglycan nanoparticles (GAGNPs) for bioprinting applications. Nanocomposite hydrogels (GLgels) were rapidly formed within seconds due to the interactions between the negatively charged groups of GAGNPs and the edges of LA. The shear-thinning behavior of the hydrogel protected encapsulated cells from aggressive shear stresses during bioprinting. The bioinks could be printed straightforwardly into shape-persistent and free-standing structures with high aspect ratios. Rheological studies demonstrated fast recovery of GLgels over multiple strain cycles. In vitro studies confirmed the ability of GLgels to support cell growth, proliferation, and spreading. In vitro osteogenic differentiation of pre-osteoblasts murine bone marrow stromal cells encapsulated inside the GLgels was also demonstrated through evaluation of ALP activity and calcium deposition. The subcutaneous implantation of the GLgel in rats confirmed its in vivo biocompatibility and biodegradability. The engineered shear-thinning hydrogel with osteoinductive characteristics can be used as a new bioink for 3D printing of constructs for bone tissue engineering applications.

Core-sheath gelatin based electrospun nanofibers for dual delivery release of biomolecules and therapeutics.

Coaxial electrospinning with the ability to use simultaneously two separate solvents provides a promising strategy for drug delivery. Nevertheless, controlled release of hydrophilic and sensitive therapeutics from slow biodegradable polymers is still challenging. To address this gap, we fabricated core-sheath fibers for dual delivery of lysozyme, as a model protein, and phenytoin sodium as a small therapeutic molecule. The sheath was processed by a gelatin solution while the core fibers were fabricated from an aqueous gelatin/PVA solution. Microstructural studies by transmission and scanning electron microscopy reveal the formation of homogeneous core-sheath nanofibers with an outer and inner diameter of 180 ± 48 nm and 106 ± 30 nm, respectively. Thermal gravimetric analysis determines that the mass loss of the core-sheath fibers fall between the mass loss values of individual sheath and core fibers. Swelling studies indicate higher water absorption of the core-sheath mat compared to the separate sheath and core membranes. In vitro drug release studies in Phosphate Buffered Saline (PBS) determine sustained release of the therapeutics from the core-sheath structure. The release trails three stages including non-Fickian diffusion at the early stage followed by the Fickian diffusion mechanism. The present study shows a useful approach to design core-sheath nanofibrous membranes with controlled and programmable drug release profiles.

Bioactive and Elastic Nanocomposites with Antimicrobial Properties for Bone Tissue Regeneration.

Bone injuries represent a major challenge in the medical field. The commonly used treatments for bone regeneration rely on the use of bone grafts that are usually associated with complications such as donor site morbidity, disease transmission, high cost, and lack of availability. Bone tissue engineering has become a golden solution for the repair of bone injuries by regenerating the damaged biological tissues using biocompatible scaffolds. However, most of the tissue engineered scaffolds do not possess the combined properties of high elasticity, appropriate stiffness, biocompatibility, osteoinductivity, and antimicrobial properties. In this study, we engineered bioactive and antimicrobial nanocomposites that can promote bone formation while simultaneously provide a barrier against bacterial infections commonly associated with bone implants. We used PEGylated polyglycerol sebacate as nanocomposites base, which was functionalized with Laponite nanosilicates, a synthetic nanoclay, and an antimicrobial peptide (AMP). The successful synthesis of the PEGylated polyglycerol sebacate and Laponite incorporation within the nanocomposites were confirmed through nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FTIR). The scaffolds had an elastic modulus and ultimate tensile strength within a range of 3.8-4.7 MPa and 1.5-3 MPa, respectively. Furthermore, the scaffolds loaded with antimicrobial peptide exhibited a significant antimicrobial activity against both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria. The in vitro cytocompatibility tests showed >90% viability of preosteoblast (W-20-17) cells. Moreover, in vitro differentiation assays demonstrated the scaffolds' ability to promote osteogenic differentiation of W-20-17. Collectively, the nanocomposites containing Laponite and antimicrobial peptide were proven to have osteoinductive and antimicrobial activity, making them desirable for bone tissue engineering applications.

Biomimetic proteoglycan nanoparticles for growth factor immobilization and delivery.

The delivery of growth factors is often challenging due to their short half-life, low stability, and rapid deactivation. In native tissues, the sulfated residual of glycosaminoglycan (GAG) polymer chains of proteoglycans immobilizes growth factors through the proteoglycans'/proteins' complexation with nanoscale organization. These biological assemblies can influence growth factor-cell surface receptor interactions, cell differentiation, cell-cell signaling, and mechanical properties of the tissues. Here, we introduce a facile procedure to prepare novel biomimetic proteoglycan nanocarriers, based on naturally derived polymers, for the immobilization and controlled release of growth factors. We developed polyelectrolyte complex nanoparticles (PCNs) as growth factor nanocarriers, which mimic the dimensions, chemical composition, and growth factor immobilization of proteoglycans in native tissues. PCNs were prepared by a polymer-polymer pair reaction method and characterized for physicochemical properties. Fourier transform infrared spectroscopy (FTIR) analysis indicated that complexation occurred through electrostatic interactions. Transmission electron microscopy (TEM) results showed that the nanocarriers had a diameter of 60 ± 11 nm and 91 ± 33 nm for dermatan sulfate sodium salt-poly-l-lysine (DS-PLL) and gum tragacanth-poly-l-lysine (GT-PLL) complexes, respectively. The colloidal nanoparticles were stable due to their negative zeta potential, i.e.-25 ± 4 mV for DS-PLL and -18 ± 3.5 mV for GT-PLL. Cytocompatibility of PCNs in contact with human bone marrow stromal cells (HS-5) was confirmed through a live/dead assay and metabolic activity measurement. In addition, vascular endothelial growth factor (VEGF) was used to evaluate the ability of PCNs to stabilize growth factors. The capability of PCNs to preserve VEGF activity for up to 21 days was confirmed by analyzing the metabolic and mitogenic characteristics of human umbilical vein endothelial cells (HUVECs). Our results demonstrated the potential applications of these nanoparticles in therapeutic delivery for tissue regeneration applications.

Anti-IL-6 eluting immunomodulatory biomaterials prolong skin allograft survival.

A primary goal in the management of burn wounds is early wound closure. The use of skin allografts represents a lifesaving strategy for severe burn patients, but their ultimate rejection limits their potential efficacy and utility. IL-6 is a major pleiotropic cytokine which critically links innate and adaptive immune responses. Here, we devised anti-IL-6 receptor eluting gelatin methacryloyl (GelMA) biomaterials (GelMA/anti-IL-6), which were implanted at the interface between the wound beds and skin allografts. Our visible light crosslinked GelMA/anti-IL-6 immunomodulatory biomaterial (IMB) demonstrated a stable kinetic release profile of anti-IL-6. In addition, the incorporation of anti-IL-6 within the GelMA hydrogel had no effect on the mechanical properties of the hydrogels. Using a highly stringent skin transplant model, the GelMA/anti-IL-6 IMB almost doubled the survival of skin allografts. The use of GelMA/anti-IL-6 IMB was far superior to systemic anti-IL-6 receptor treatment in prolonging skin allograft survival. As compared to the untreated control group, skin from the GelMA/anti-IL-6 IMB group contained significantly fewer alloreactive T cells and macrophages. Interestingly, the environmental milieu of the draining lymph nodes (DLNs) of the mice implanted with the GelMA/anti-IL-6 IMB was also considerably less pro-inflammatory. The percentage of CD4+ IFNγ+ cells was much lower in the DLNs of the GelMA/anti-IL-6 IMB group in comparison to the GelMA group. These data highlight the importance of localized immune delivery in prolonging skin allograft survival and its potential utility in treating patients with severe burns.